LOCUS       sequence-464290-        6836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid for integration of target sequence at HO locus, Kan 
            selection, pGAL inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6836)
  AUTHORS   Voth WP, Richards JD, Shaw JM, Stillman DJ
  TITLE     Yeast vectors for integration at the HO locus.
  JOURNAL   Nucleic Acids Res. 2001 Jun 15;29(12):E59-9.
  PUBMED    11410682
REFERENCE   2  (bases 1 to 6836)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2001-06-15"; volume: "29(12)"; pages: "E59-9"
COMMENT     Sequence Label: HO-pGAL-poly-KanMX4-HO
FEATURES             Location/Qualifiers
     source          1..6836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..248)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     214..232
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     250..266
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     251..267
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(301..317)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(332..773)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(338..361)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    656..773
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     gene            846..2202
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        846..1189
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1190..1999
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1257..1276)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2005..2202
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2810..2829)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2811..2829)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2836..2853)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2836..2852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2845..2867)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3152..3171)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3284..3305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3459..3478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3578..3600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3638..3656)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3724..3828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3829..3897,3898..4689)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4047..4066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4860..5448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5349..5368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5602..5619
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5736..5757
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    5812..5828
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5817..5839
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5836..5852
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5836..5852
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcaaattaaa gccttcgagc gtcccaaaac cttctcaagc aaggttttca gtataatgtt
       61 acatgcgtac acgcgtctgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct
      121 taatactaac ataactataa aaaaataaat agggacctag acttcaggtt gtctaactcc
      181 ttccttttcg gttagagcgg atgtgggggg agggcgtgaa tgtaagcgtg acataactaa
      241 ttacatgact cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
      301 gatccactag ttctagaatc cggggttttt tctccttgac gttaaagtat agaggtatat
      361 taacaatttt ttgttgatac ttttattaca tttgaataag aagtaataca aaccgaaaat
      421 gttgaaagta ttagttaaag tggttatgca gtttttgcat ttatatatct gttaatagat
      481 caaaaatcat cgcttcgctg attaattacc ccagaaataa ggctaaaaaa ctaatcgcat
      541 tatcatccta tggttgttaa tttgattcgt tcatttgaag gtttgtgggg ccaggttact
      601 gccaattttt cctcttcata accataaaag ctagtattgt agaatcttta ttgttcggag
      661 cagtgcggcg cgaggcacat ctgcgtttca ggaacgcgac cggtgaagac gaggacgcac
      721 ggaggagagt cttccttcgg agggctgtca cccgctcggc ggcttctaat ccgtactaga
      781 ggggttaatt aaggcgcgcc agatctgttt agcttgcctc gtccccgccg ggtcacccgg
      841 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
      901 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
      961 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
     1021 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     1081 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     1141 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tgggtaagga
     1201 aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1261 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     1321 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     1381 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     1441 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gcaaaacagc
     1501 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     1561 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     1621 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     1681 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
     1741 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     1801 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     1861 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     1921 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     1981 gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     2041 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     2101 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     2161 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     2221 cgccgccatc cagtgtcgaa aacgagctcg aattaattcc tgggggaaca acttcacaga
     2281 atgttttgtc atattgtcga agtggtcaca aaacaagaga agttccgcca attataaaaa
     2341 gggaacccgt atatttcagc ttcacggatg atttccaggg tgagagtact gtatatgggc
     2401 ttacgataga aggccataaa aatttcttgc ttggcaacaa aatagaagtg aaatcatgtc
     2461 gaggctgctg tgtgggagaa cagcataaaa tatcacaaaa aaagaatcta aaacactgtg
     2521 ttgcttgtcc cagaaaggga atcaagtatt tttataaaga ttggagtggt aaaaatcgag
     2581 tatgtgctag atgctatgga agatacaaat tcagcggtca tcactgtata aattgcaagt
     2641 atgtaccaga agcacgtgaa gtgaaaaagg caaaagacaa aggcgaaaaa ttgggcatta
     2701 cgcccgaagg tttgccagtt aaaggaccag agtgtataaa atgtggcgga atcttacagt
     2761 ggcctatgcg gccgctctag ctagaactag tggatcgatc cccaattcgc cctatagtga
     2821 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2881 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2941 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     3001 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     3061 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3121 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     3181 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gtgggccatc gccctgatag
     3241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3301 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     3361 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3421 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     3481 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3541 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3601 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3661 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3721 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3781 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3841 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3901 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3961 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4021 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4081 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4141 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4201 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4261 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4321 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4381 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4441 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4501 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4561 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4621 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4681 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4741 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4801 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4861 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4921 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4981 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5041 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5101 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5161 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5221 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5281 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5341 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5401 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5461 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5521 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5581 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     5641 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     5701 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     5761 gcaccccagg ctttacactt tatgcttccg cggctcgtat gttgtgtgga attgtgagcg
     5821 gataacaatt tcacacagga aacagctatg accatgatta cgccactagt ccgaggcctc
     5881 gagatccgat atcgccgtgg cggccgccag ctgaagctta attatcctgg gcacgagtga
     5941 aacaaagcta aaacctttat ttagcatggc cattgaatgt aacaattata tatatcgcaa
     6001 gcacaaaaaa tcaaggagag agaactacca ctttgttcat gtgtacaatg ttcattatct
     6061 ccataagcaa aaaaaaaaaa atagaaaaca tatgctataa ggttgatatt ctcacgagta
     6121 agcggcactt gctacttatt gacattgcag atttttggct acagaaatag tatattagag
     6181 attataattg ctaatcaaat caaaatataa aattagtaaa ccaaaccatt tatacccttc
     6241 cttagtagtt atggattgtt ttttaatgat atttctgcaa accaaagaaa gattgttatc
     6301 cagatagaat ttagttttga tattcatttt tttgttgaag attgaacgcc atatctgggc
     6361 ctcataattc aaaagacggt gccattatcg gtagcgtttc gcattgtact ggatttcaga
     6421 aatttcacag ttgatgaatc gaaaagaatg gtctcattgc aacacgtaag gttaagatgt
     6481 ccctttttac cattataggc aataaatgaa tcataaaacg accgtatact ggtgaaatag
     6541 tagggagaac gagtacctgt agtaaaaagt ataaatcata gttaatcggg caatgtccct
     6601 cgatcaagga gtattgtgtc atgttcgaga caaacgccaa catttttgtt tcttttggac
     6661 aaatgttgtt tgcatttatg atccgttata ttttgatcta atgtagagtt gcacgtagtt
     6721 cttactggca aagaaatcga tgcataccaa aaaagaataa aggtgatatt tgatctttac
     6781 cgtttagttc caacgtaaaa ttgtgccttt ggacttaaaa tggcgtcgta ccggcc
//
